pTCF176 HLA-A*02:01 (CD8 mut)
PurposeE. coli expression of soluble MHC protein
|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||180459||Standard format: Plasmid sent in bacteria as agar stab||1||$85|
This material is available to academics and nonprofits only.
Vector typeBacterial Expression
Growth in Bacteria
Bacterial Resistance(s)Kanamycin, 50 μg/mL
Copy numberLow Copy
Gene/Insert nameHLA-A*02:01 (CD8 mut)
SpeciesH. sapiens (human)
MutationD227K, T228A, replacement of transmembrane domain with enzymatic biotinylation site
/ Fusion Protein
- BSP41 (C terminal on insert)
- Cloning method Restriction Enzyme
- 5′ cloning site Unknown (unknown if destroyed)
Terms and Licenses
- Not Available to Industry
- Zeocin® is an InvivoGen trademark.
Mutation prevents binding of CD8 (Purbhoo et al. (2001) J. Biol. Chem. doi.org/10.1074/jbc.M102498200). See https://www.ebi.ac.uk/ipd/imgt/hla/alleles/allele/?accession=HLA00005 for allele sequence data. For detailed protocols of expression, purification and use to visualize antigen-specific T cells, please refer to Altman and Davis (2016) Curr. Protoc. Immunol. doi.org/10.1002/cpim.14. We recommend the E. coli BL21(DE3) strain for expression.
These plasmids were created by your colleagues. Please acknowledge the Principal Investigator, cite the article in which the plasmids were described, and include Addgene in the Materials and Methods of your future publications.
For your Materials & Methods section:pTCF176 HLA-A*02:01 (CD8 mut) was a gift from John Altman (Addgene plasmid # 180459 ; http://n2t.net/addgene:180459 ; RRID:Addgene_180459)